Method of preparing 3alpha-acetoxy-17alpha-hydroxy etiocholane-11-one



United States Patent METHOD OF PREPARING Sa-A'CETOXY-fla-HY- .DROXY .ETIOCHOLANE-ll-ONE JulieuWarnant, Neuilly-sur-Seine, and Robert Joly, Monitmorency, France, assignors to Les Laboratoires Franmis de Chimiotherapie, Paris, France, a French body corporate 1 No Drawing. Application February 11, 1958 Serial No. 714,470

Claims priority, application France February 12, 1957 2 Claims. (Cl. 260-39745) This invention relates to trisubstituted derivatives or etiocholane and to a method of preparing same.

More particularly, the present invention relates to a method of preparing Ba-acetoxy 17-hydroxy etiocholane ll-one in two stereoisomeric forms, 3a-acetoxy .17a-hydroxy etiocholane ll-one, II, and Sa-acetoxy 17 fl-hydroxy etiocholane ll-one III. The Romannumerals indicated after the chemical names represent the formulas hereinaftergiven.

These compounds have interesting physiological and pharmacodynamic properties, and may also serve asintermediate products in the preparation of the corresponding .1 111- or 1 lfi-hydroxyl-ated derivatives.

, The process according to the invention consists in de-' composing the ,20-dihydroperoxide of 3a-acetoxy pregnane 11,20-dione, I, by the action of either heat or ultraviolet rays in the presence of a solvent such as methanol, ethanol and the like or by combining the .action of heat with the action of the ultraviolet rays in order to accelerate the reaction. a

The following equation illustrates the reaction of the method herein a In United States .patent .application aentitledfNew Derivatives of Ketosteroids and Method of Preparing Same filed June 6, 1957, Serial No. 663,880, now Patent No. 2,842,544, the applicants have described the preparation of the ZO-dihydroperoxide of 3a-acetoxy pregnane'111,20- ,dione, I, by the action of anhydroushydiogeri peroxide in an organic solvent on 3a-acetoxy pregnane 11,20- dione.

It has now been found that, in the presence of certain solvents such as methanol, or ethanol and the like, the action of heat or ultraviolet rays causes decomposition of the dihydroperoxide, said decomposition becoming more rapid as one approaches the boiling point of the 2,927,120 Patented Mar. 1, 1960 "ice solvent used, the process being accelerated by the simul- ,taneous supply of the two forms of energy. Theproduets which 01m, in the course of the reaction are, on the one hand, mainly 3u-acetoxy pregnane 11,20-dione which can again be converted into the starting dihydroperoxide, on theother ,hand,:the reaction produces an acid fraction and a mixture of 3a-acetoxy l'la-hydroxy etiocholane .11-.one, .11, of 3aacetoxy l7,B-hydroxy etiocholane 1.1- one, III, and of 3a-acetoxy etiocholane ll-one. These last three compounds can be separated easily by chromatography.

The reaction may be followed by the dosage of the active oxygen of the dihydroperoxide introduced. Thereaction is terminated when the same has disappeared.

With the above featuresin mind, it is an object of the present invention to provide a method of preparing derivatives of etiocholane.

Another object of the present invention is to prepare stereoisomers of 30: acetoxy 17-hydroxy etiocholane 11-one.

A further object of the present invention is to prepare stereoisomers of Swacetoxy 17 hydroxy etiocholane ll-one having physiological and pharmacodynamic properties and which are useful in the preparation of their corresponding 11a or 1113 hydroxylated derivatives.

A specific object of the present invention is to provide 3a acetoxy 17a hydroxy etiocholane ll-one.

Another specific object of this invention is to provide 30: 'acetoxy 17,3 'hydroxy etiocholane l'l-one.

With the above and other objects and features in view, the preferred method of carrying out the process of the present invention, consequent-1y, consists in causing a mixture of dihydroperoxide, I, and methanol or ethanol to boil while under the influence of ultraviolet irradiation until compound I has disappeared. On to cooling, a portion of the 3a-acetoxy pregnane 11,20-dione that has Jon M ,formed, crystallizes. .After evaporation to dryness, the

mother liquors of crystallization are taken up with aneutral solvent which is immiscible with water, and then freed the acid fraction :thathas formed by washing a mild alkaline substance such as sodium bicarbonate. The resulting neutral fraction obtained is then subjected to treatment with the reagent T of Girard,

. whichmakes it possible to recover another fraction of of methanol.

' invention without however limiting its scope, it is possible to change the nature of the solvent and the temperature,

to heat without irradiating or to irradiate without heating, or to carry out at'once a separation by chromatography of the reaction mixture without thereby exceeding the scope of the invention. In the description here- 'in given, melting points are instantaneous melting points determined on the Maquenne block.

EXAMPLE Prepart ztion of 3a-acetoxy 17-hydr0xy etiocholane 11- 'one in its two stereoisomeric frms, F0rmulas II and Ill 50 g. of dihydroperoxide, I, are suspended in 1500 cc. While irradiating the suspension such as by means of two ultraviolet Hanau 5.80 lamps and stirring the suspension mechanically, it is heated to boiling under reflux, until starch-iodide paper applied for testing no longer reveals the presence of active oxygen. This treatment takes about 9 hours. The solution is then cooled and concentrated in vacuo at a low temperature -to about one seventh of its starting volume, and then cooled to -10 C. Crystallization is allowed to proceed for about one hour, and the crystals that have formed are separated by filtration or centrifugation and washed with methanol and then dried.

This method produces g. of 3a-acetoxy pregnane 11, ZO-dione. After the mother liquors of-crystallization have been evaporated to dryness, the residue is then dissolved in ether and the acid fraction is eliminated by washing with an aqueous solution saturated with sodium bicarbonate or other suitable mild alkaline agent. The ethereal extract is then washed with water, then dried -over magnesium sulfate and evaporated to dryness in vacuo. The residue thus obtained is treated, underthe usual conditions, with an equal weight of the Girard T reagent. This produces, on the one hand, an aqueous solution of T hydrazones which, by acidification, again yields 10 g. of Ba-acetoxy pregnane 11,20-dione bringing the total of the compound recovered or produced to 50% by weight of the initial dihydroperoxide, and, on the other hand, an ethereal extract comprising the products which have not combined with the T reagent. The neutral ethereal extract is evaporated to dryness and then taken up with 600 cc. of cyclohexane. Filtering is effected to eliminate an insoluble product or solid and chromatography is carried out over alumina.

The alumina is then eluted with benzene, then hen-.-

zene-ether mixtures with the following ether contents 1%,2%,5%, l0%,20% and 50%.

The benzene eluate yields, after evaporation of the solvent and recrystallization in methanol, Bot-acetoxy etiocholane 11 one, having a M. P. 130 C., [a] =+74 C. (c.=0.5%, acetone), which has previously been described in the French Patent No. 1,173,938.

The first benzene-ether eluates containing up to 10% of ether produce, after evaporation to dryness, 2 g. of Six-acetoxy 17oc-l'lYdl'0XY etiocholane ll-one, II.. After recrystallization in methanol, there is obtained a product having a M.P.=172 C., [a] =+8O 0:3 (c.= 0.5% acetone). This product occurs in the form of colorless needles, is slightly soluble in acetone and alcohol, insoluble in ether and Water. effect.

I h s a de n te" d o nic' C H O and a molecular weight of 348.47. As calculated: C=72.38%, H=9.26%,O=18.37%. As found: C=72.1%, H=9.2%, O=18.6%.

This compound has not been described in the past.

By chromic acid oxidation it yields 3a-acetoxy etiocholane 11,17-dione, M.P. 162 C., which is known in the art.

The benzene-ether washings from 20% to 50% of ether yield, by evaporation to dryness, 3a-acetoxy 1713-hydroxy etiocholane ll-one, III, having a M.P. C., which is new and has not been described in literature heretofore, and which also yields, by chromic acid oxidation, 3w acetoxy etiocholane 11,17-dione, M.P. 162 C.

Its analysis shows the formula C H O with a molecular Weight of 348.47. As calculated: C=72.38%, .H=9.26%, O=18.37%. As found: C=72.4%,

From the foregoing description taken in conjunction with the accompanying illustrative example, it will be noted that there is provided a new method of forming new derivatives of etiocholane which have valuable uses in physiology, pharmacodynamics and biochemistry.

While a preferred method and new products have been 'described, it is to be understood that changes as to steps ,or procedure or use of materials may be made without departing from the spirit and scope of the invention as claimed.

We claim:

1. In a process of preparing 3m-acetoxy-17-hydroxy etiocholane-l l-one in its stereo-isomeric forms, the steps which comprise heating to boiling under reflux 'a suspension of the dihydroperoxide of 3a-acetoxy-pregnane- 11,20-dione in a lower alkanol while exposing the suspension to the action of ultraviolet rays, until the di- 'hydroperoxide has disappeared therefrom, cooling the reaction mixture to cause crystallization of Zia-acetoxy pregnane-11,20-dione, separating said crystallized 3aacetoxy pregnane-11,20-dione from the mother liquor, removing acid by-products from the mother liquor by washing with an alkali metal bicarbonate solution, reacting the washed mother liquor with Girards reagent T to cause formation of the Girards hydrazone T with 30:- acetoxy pregnane-11,20-dione, separating said hydrazone from the unreacted components of the mother liquor, subjecting a solution of said unreacted components in cyclohexane to a chromatographic treatment over alumina, eluting the alumina successively with benzene and mixtures of benzene and progressively increasing amounts of ether, and isolating 3a-acetoxy-l7a-hydroxy etiochw lane-1 l-one and 3a-acetoxy-17fi-hydroxy etiocholane-llone by evaporation of the benzene-ether eluates.

2. The process according to claim 1, wherein 3a-acetoxy-l7whydroxy etiocholane-ll-one is isolated from the eluates obtained by eluting with benzene-ether mixtures containing up to 10% of ether.

References Cited in the file of this patent OTHER REFERENCES Fieser: Natural Products Related to Phenanthrene. 1949,page 402. V i 

1. IN A PROCESS OF PREPARING 3A-ACETOXY-17-HYDROXY ETIOCHOLANE-11-ONE IN ITS STEREO-ISOMERIC FORMS, THE STEPS WHICH COMPRISE HEATING TO BOILING UNDER REFLUX A SUSPENSION OF THE DIHYDROPEROXIDE OF 3A-ACETOXY-PREGANANE11,20-DIONE IN A LOWER ALKANOL WHILE EXPOSING THE SUSPENSION TO THE ACTION OF ULTRAVIOLET RAYS, UNTIL THE DIHYDROPEROXIDE HAS DISAPPEARED THEREFROM, COOLING THE REACTION MIXTURE TO CAUSE CRYSTALLIZATION OF 3A-ACETOXY JPREGNANE-11,20-DIONE, SEPARATING SAID CRYSTALLIZED 3AACETOXY PREGNANE-11,20-DIONE FROM THE MOTHER LIQUOR, REMOVING ACID BY-PRODUCTS FROM THE MOTHER LIQUOR BY WASHING WITH AN ALKALI METAL BICARBONATE SOLUTION, REACTING THE WASHED MOTHER LIQUOR WITH GIRARD''S REAGENT T TO CAUSE FORMATION OF THE GIRARD''S HYDRAZONE T WITH 3AACETOXY PREGNANE-11,20-DIONE, SEPARATING SAID HYDRAZONE FROM THE UNREACTED COMPONENTS OF THE MOTHER LIQUOR, SUBJECTING A SOLUTION OF SAID UNREACTED COMPONENTS IN CYCLOHEXANE TO A CHROMATOGRAPHIC TREATMENT OVER ALUMINA, ELUTING THE ALUMINA SUCCESSIVELY WITH BENZENE AND MIXTURES OF BENZENNE AND PROGRESSIVELY INCREASING AMOUNTS OF ETHER, AND ISOLATING 3A-ACETOXY-17A-HYDROXY ETIOCHOLANE-11-ONE AND 3A-ACETOXY-17B-HYDROXY ETIOCHOLANE-11ONE BY EVAPORATION OF THE BENZENE-ETHER ELUATES. 